Influence of enzyme solution on protoplast culture and transient gene expression in maize (Zea maysL.)
✍ Scribed by Brigitte Krautwig; Paul A. Lazzeri; Horst Lörz
- Publisher
- Springer
- Year
- 1994
- Tongue
- English
- Weight
- 783 KB
- Volume
- 39
- Category
- Article
- ISSN
- 0167-6857
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✦ Synopsis
The influence of two enzyme solutions, differing only in the presence or absence of Macerozyme, on protoplast yield, colony formation and transient GUS (/~-glucuronidase) activity was studied. For all parameters tested the presence of Macerozyme during protoplast isolation had a negative influence. Using an enzyme solution without Macerozyme suspension aggregates gave up to 4.4 times higher protoplast yield and plating efficiencies were increased up to 10-fold. Further, protoplasts isolated without Macerozyme showed a 5.2-fold higher GUS activity in transient gene expression. Apart from the presence of Macerozyme, longer incubation (3 compared with 1.5 h) of cell aggregates in the enzyme solution also had a negative effect on transient transformation efficiency. These data demonstrate that protoplast isolation conditions have a profound effect on transient gene expression and it is proposed that these factors will also influence stable transformation efficiency.